Lung cancer remains the leading cause of cancer-related deaths worldwide. Surgical removal of the tumor is the best therapeutic option for patients with non-small cell lung cancer, yet up to 20% of patients still leave the operating room with deposits of cancer left behind. Advances in CAT and PET scans have provided surgeons with helpful radiographic imaging information prior to surgery, but have limited specificity for nodules less than 5-7mm in diameter. Subsequent pathological analysis confirms/refutes what a surgeon has resected. However, both of these procedures require additional time and occur outside of the operating room, providing little real- time support during the surgery. The applicant has developed a novel cocktail that combines two ligands targeted specifically to lung tumor tissue tagged with near infrared (NIR) dyes. The fluorescent cocktail containing ligands targeted to both folate receptor and the CCK2 receptor will essentially identify tumor populations that overexpress either or both of these cell surface receptors. Real-time visualization not only of the complete tumor margins but also potentially other malignant sites outside the primary tumor area will enable more complete surgical removal of primary and other tumor tissue. Moreover, fluorescence imaging could aid in the surgical decision to conserve lung function by removing only the diseased tumor tissue rather than lobectomy or pneumonectomy. The significance of this tumor-targeted fluorescent cocktail is that it will provide real-time visualization of tumor tissue during surgery - a valuable tool to differentiate tumor from healthy tissue. In this Phase I SBIR proposal, the applicant will demonstrate the feasibility of co-stainin with the cocktail in cell culture and human lung carcinoma tissue samples. Studies in mice implanted with tumor cells will demonstrate specificity and sensitivity of the levels of detection n vivo. Our commercialization efforts will be aimed at the development of the cocktail for intraoperative use in lung cancer resection surgery. The immediate objective of this work is to provide proof of concept for the NIR-ligand cocktail both in vitro and in vivo. The ultimate goal i to introduce a commercially viable product that will enable more complete tumor removal and a reduction in the recurrence rate of pulmonary cancer and its associated morbidity, mortality, and expense.